Motoyoshi Maruno

Association of human papillomavirus type 6 with a verruciform xanthoma.

We report a case of verruciform xanthoma (VX) associated with human papillomavirus (HPV) in a 67-year-old male. The patient had a pale-reddish, granular and verrucous tumor on the right side of his scrotum for four years. Histopathologic examination showed typical features of VX. HPV was detected by immunohistochemistry, electron microscopy, and PCR examinations. Ultrastructural examination revealed virus-like particles of 40–50 nm in the nucleus of the upper epidermal keratinocytes. HPV type 6a DNA was detected in lesional tissue by polymerase chain reaction and sequence analysis. To the best of our knowledge, this is the first case report of VX associated with HPV.

Comparative Studies of the Detection Rates of Leishmania Parasites from Formalin, Ethanol‐fixed, Frozen Human Skin Specimens by Polymerase Chain Reaction and Southern Blotting

In this study, detection rates of Leishmania parasites from human skin were compared among three different types of specimens, formalin‐fixed, ethanol‐fixed, and frozen, by polymerase chain reaction (PCR) and Southern blotting. For this purpose, we used biopsy specimens collected from 19 leishmaniasis patients and performed PCR and Southern hybridization with the probe specific for Leishmania (Viannia) braziliensis complex. Among these 19, 16 specimens were from cutaneous leishmaniasis (CL), one, diffuse cutaneous leishmaniasis (DCL) and 2, mucocutaneous leishmaniasis (MCL) and were formalin‐fixed and paraffin‐embedded. The causative agents for one case of CL and one case of DCL were already identified as L. (Leishmania) complex. Six specimens of CL were preserved in 100% ethanol. Two specimens of MCL were frozen tissues. PCR using the formalin‐fixed and paraffin‐embedded specimens revealed positive bands at 70bp in 9 (47.4%) out of 19 specimens of CL, MCL and DCL. Southern blotting detected the signals in 12 (63.2%) out of the 19. PCR using the 100% ethanol‐fixed specimens revealed positive bands in 4 (66.7%) out of 6, and Southern blotting also detected the signals in 4 (66.7%) out of the 6. PCR and Southern blotting using 2 frozen specimens of MCL were always positive (100%). Although we failed to detect significant differences by Chi‐square test between the results from the formalin‐fixed, paraffin‐embedded specimens and those from 100% ethanol‐fixed ones, we concluded that ethanol‐fixed specimens, convenient for transportation and storage, would be more useful for diagnosis of leishmaniasis by PCR in a developing country.

Low-dose UVB contributes to host resistance against Leishmania amazonensis infection in mice through induction of gamma interferon and tumor necrosis factor alpha cytokines.

ABSTRACT UV radiation suppresses the immune response, a fact which raises the question of whether the phenomenon may find practical applications in the outcome of infectious diseases. In this study, BALB/c mice were exposed to low-dose UVB (250 J/m2) from Dermaray M-DMR-100 for 4 consecutive days. Twelve hours after the last UV exposure, groups of mice were injected with 2 × 106Leishmania amazonensis promastigotes. The development of skin lesions, as assessed by measurement of visible cutaneous lesions, was significantly suppressed in low-dose UVB-irradiated mice compared to nonirradiated controls. In order to characterize the cytokines involved in this phenomenon, BALB/c mice were irradiated with identical doses of UVB, and gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin 4 cytokine levels in blood serum and skin were examined at different times by a sandwich enzyme-linked immunosorbent assay, immunohistochemical analysis, and reverse transcription (RT)-PCR. Upregulated expression of serum IFN-γ and TNF-α was observed from 6 to 24 h. Positive results for IFN-γ and TNF-α in UVB-irradiated mice were obtained by immunohistochemical analysis. By RT-PCR, the mRNA expression of both IFN-γ and TNF-α cytokines was detected in a time-dependent manner only in UVB-irradiated mice. Histopathological analysis and electron microscopy revealed that cellular infiltration, tissue parasitism, and parasitophorus vacuoles in irradiated mice were markedly less noticeable than those in nonirradiated controls. These results suggested that low-dose UVB irradiation played a pathogen-suppressing role in Leishmania-susceptible BALB/c mice via systemic and local upregulation of Th1 (IFN-γ and TNF-α) cytokines.

Pre-exposure with low-dose UVA suppresses lesion development and enhances Th1 response in BALB/c mice infected with Leishmania (Leishmania) amazonensis

This study was conducted to determine whether exposing mice to ultraviolet (UV) radiation would alter the pathogenesis of infection with Leishmania (Leishmania) amazonensis (L. amazonensis) which causes progressive cutaneous disease in susceptible mouse strains. BALB/c mice were irradiated with 10 and 30 J/cm(2) UVA on shaved skin of the back from Dermaray (M-DMR-100) for 4 consecutive days before infection with Leishmania promastigotes. The course of disease was recorded by measuring the size of lesions at various times after infection. Mice groups irradiated with UVA 10 and 30 J/cm(2) showed significantly suppressed lesion development compared with the non-irradiated mice. Light and electron microscopy revealed a few parasites at the site of inoculation in UVA-irradiated subjects. Sandwich enzyme-linked-immunosorbent-assay (ELISA) examination of sera showed dose dependently upregulated interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12, and downregulated interleukin (IL)-4 and interleukin (IL)-10 levels in UVA-irradiated as compared with the non-irradiated mice. Positive signals for IFN-gamma mRNA in irradiated mice were obtained by RT-PCR, while non-irradiated mice showed negative results. None of the examined samples showed signal for IL-4 mRNA. The present study disclosed that exposure of mice to different low-doses of UVA irradiation prior to infection may interfere with immunity to L. amazonensis in the murine model. This indicates that the cell-mediated response switch from Th2 to Th1 pattern suppressed the cutaneous lesions of L. amazonensis.

An assessment of the malignant potential of actinic keratoses and Bowen's disease : p53 and PCNA expression pattern correlate with the number of desmosomes

Actinic keratoses (AK) and Bowens disease (BD), both intraepidermal skin tumors, have a potential progression to squamous cell carcinoma (SCC). To evaluate the malignant potential of AK and BD, the expression pattern of p53 protein and proliferating cell nuclear antigen (PCNA) were examined in five types of AK and BD by immunohistochemistry. The ultrastructural difference of epidermal cells between AK and BD lesions was investigated. In the study of p53 and PCNA expression, the atrophic and acantholytic types of AK showed lower positive rates compared to others. These two types did not demonstrate all layers expression pattern. The number of desmosomes of the epidermal cells was significantly reduced in BD, and in the bowenoid and hypertrophic types of AK compared with other types of AK. The number of hemi‐desmosomes showed greatest reduction in BD and the bowenoid type of AK. On the basis of our findings, it is hypothesized that atrophic and acantholytic types of AK may have the lowest, and the bowenoid type of AK and BD may have the highest, malignant potential.

Amyloid deposition is frequently observed in skin lesions of hypertrophic lupus erythematosus.

Four cases of Hypertrophic Lupus Erythematosus (HLE) were reported. The lesions of HLE were observed on the forearms, face and hands in all four cases. Clinically, the lesions were erythematous, hyperkeratotic plaques. The clinical course was marked by chronicity and progression of the lesion. Histologically, marked hyperkeratosis, parakeratosis, acanthosis, degenerative changes of basal cells in H/E stain, and thickened, multilayered basement membrane in PAS stain, were observed. The observations of Dylon stain revealed that localized amyloid deposition was observed in all four cases of HLE lesions, as fluorescent‐orange colored amyloid deposits in the papillary dermis and subepidermal areas at near or just below the dermo‐epidermal junction appeared under fluorescent microscope. On the basis of clinical and histological observations, we suggest that chronic irritation, such as sunlight exposure over a long‐duration, might have caused the characteristic abnormalities at the dermo‐epidermal junction and also initiated the frequency of amyloid deposits locally secondary to the diseases. We compared our HLE cases to other types of lupus erythematosus (LE) skin lesions, as to whether deposition of amyloid materials were frequently observed or not. Amyloid deposition was observed in one case of DLE and none of the SLE cases. Localized amyloid deposition was more frequently observed in skin lesions, secondary to HLE disease, as compared to other types of LE.

Molluscum Contagiosum Appearing as a Solitary Lesion on the Eyelid

To the Editor: Molluscum contagiosum (MC) is an infectious disease of the skin and mucous membranes caused by a DNA virus belonging to the pox group of viruses. It was first reported by Bateman et al. as a clinical entity with a contagious nature in 1814 (1). MC is generally a self-limiting, benign skin disease. In children, infection is spread by direct contact or through the fomites, with lesions predominating on the trunk, extremities, and face. In young adults, this infection is spread primarily through sexual contact, and lesions develop on the lower abdomen, thighs, buttocks, and genitalia. Typical lesions are pale, oval and/or round, raised papules with an umblicated center and usually widespread distribution. The disease can be sporadic or endemic; poverty, overcrowding, moist environments, and poor hygiene facilitate its spread (2). Involvement of the eyelids as a solitary lesion in a healthy adult has rarely been reported. We experienced an atypical case of MC appearing as a solitary eyelid lesion on a healthy adult patient. Case Report: A 55-year-old male visited our outpatient clinic with the complaint of a small vesicle near the right eyelid, which had slowly increased in size and became granulomatous. He had no pain or itching. None of his family members suffered from the same complaint. On physical examination, the lesion was a 4 x 3 mm-sized, oval, firm, well circumscribed and erythematous papule near the right eyelid (Fig. 1). No immunodeficient conditions had been diagnosed. The patient was treated by surgical excision of the lesion; up to date no recurreneehad.been observed. Light microscopic examination showed invagination of keratotic column with an onion-shaped acantholytic epidermis containing basopilic granular inclusion bodies Fig. 1. Clinical photograph of the patient showing a solitary lesion on the right lower eyelid

A Case of Trichorrhexis Nodosa Developed in Winter

A young Japanese man developed localized trichorrhexis nodosa (LTN) of the scalp hair in the winter season. To investigate the roles of shampoo, severe sunlight exposure and/or mechanical injuries, we performed the following studies. Hair was collected from the patient and from a control. The study was performed in two steps. In the first step, hair was put into shampoo, rinsed with saline water, and then exposed to ultraviolet B (UVB) radiation once a day for one week. In the second step, the hair was similarly treated, but each shaft was bent gently with forceps before UVB exposure. Scanning electron microscopic studies revealed cuticular changes when the hair was treated only with shampoo and UVB. When it was treated with shampoo, UVB, and mechanical bending, the patients hair developed longitudinal and transverse fractures of the hair shafts, while the control hair showed only partial damage to the hair shaft. On the basis on the above findings, we conclude that mechanical bending may damage the hair shaft.

Detection of species of the subgenus Leishmania parasites using polymerase chain reaction and southern blotting.

In this study, an attempt was made to identify different Leishmania species by polymerase chain reaction (PCR). Fourteen Leishmania strains from stock were tested by PCR and Southern blotting. A pair of primers were employed that anneal to the kinetoplast DNA sequence conserved among subgenus Leishmania. Of the 14 Leishmania strains used in this study, six showed strong bands of approximately 170 bp, and all the positive strains belonged to the species of the subgenus Leishmania viz., Leishmania (Leishmania) garnhami, L. (L.) amazonensis, L. (L.) pifanoi, L. (L.) mexicana, L. (L.) chagasi, and L. (L.) major. All the species belonging to the subgenus Viannia used in this study were negative by PCR. These results suggest that the primer pair may be useful for identification of the species belonging to the subgenus Leishmania of the New World as well as to distinguish subgenus Leishmania from subgenus Viannia.